血清HCV RNA不同提取方法的对比研究

COMPARATIVE STUDY AMONG DIFFERENT METHODS FOR EXTRACTING HCV RNA FROM SERUM

目的:探讨不同方法提取丙型肝炎患者血清HCVRNA,比较每种方法提取的RNA用于RT-PCR荧光定量检测的敏感度。方法:分别采用异硫氰酸胍-酚氯仿法、蛋白酶K-酚氯仿法、固相吸附法、柱式总RNA抽提法提取7例疑似HCV感染者血清标本中的HCVRNA,采用实时荧光定量PCR法进行检测。结果:采用异硫氰酸胍-酚氯仿法、蛋白酶K-酚氯仿法提取HCVRNA用于核酸检测有漏检现象,而采用固相吸附法和柱式总RNA抽提法,敏感度均可达到100%。结论采用固相吸附法和柱式总RNA抽提法提取血清HCVRNA具有较高的敏感度,在实时荧光定量PCR法检测中具有重要的临床价值。

OBJECTIVE ： To study the different methods to extract HCV RNA from serum of hepatitis C patients and compare the sensitivity of each method for RT - PCR quantitative fluorescence detection. METHOD ： HCV RNA was extracted from 7 cases with suspected HCV infection by method of acid Gtmidinlum Thioeyanate - phenol - choroform extracton, protease - K - phenol- choroform extraction, solid- phase extracton and the 3S column extraction. Real -time quantitative PCR method was used for detection. RESULTS ： There were one or two missed for the method of Thioeyanate - phenol - choroform extracton and protease- K- phenol- choroform extraction, while solid- phase extraction and the 3S column extraction was 100% sensitive. CONCLUSION： using solid - phase extraction and the 3S column extraction to extract HCV RNA in serum can obtain a high sensitivity for real - time quantitative PCR , which has an important value in clinical practice.